What chemical modification does Synthego offer? What are the advantages?

For a chemically modified version of our CRISPRevolution products, we offer 2’-O-methyl analogs and 3’ phosphorothioate internucleotide linkages at the first three 5’ and 3’ terminal RNA residues in the following configuration:

sgRNA: 5′-U*A*A*UUUCACAGCUGCACAUA+Synthego Scaffold…U*U*U*-3′

OR

crRNA: 5′-U*A*A*UUUCACAGCUGCACAUA+Synthego Linker…U*G*G*-3′

+

tracrRNA: 5′-A*A*A*+Synthego Scaffold…U*U*U*-3′

*=modification

Chemicalmodifications provide greater stability and protection from exonucleases to guide RNAs, allowing them to persist within cells for longer than unmodified guide RNAs. This persistence likely allows Ribonucleoprotein (RNP) complexes containing guide RNAs to access regions of the genome that may be challenging for Cas9 to interrogate due to cell cycle or heterochromatin bunching factors. In many primary cells, stem cells and cell lines of hematopoietic origin, chemicallymodified sgRNAs also protect against innate intracellular immune responses that can actively degrade RNA or trigger immune cascades that lead to cell death.

Note that: The standard chemical modifications provided by Synthego are not suited for use with Cpf1-based gRNA. Please contact us if you are interested in specialty modifications that are currently being evaluated for use with Cpf1.

If you’re interested in a chemically modified option, select Modifications from the dropdown menu when inputting your target sequence on the order page. If you would like alternative configurations or modifiers, please contact us.

Click here for more information about chemically modified synthetic gRNA.